The molecular events associated with the development of renal cell carcinoma are essentially unknown and useful biochemical markers for this tumor are not available. High-resolution protein separation techniques have been used in preliminary experiments to identify a group of proteins which are associated with the tumor, but absent or significantly reduced in normal renal cortex. This application has as its long-term objective the understanding of the role these proteins play in the expression of the malignant phenotype and the determination of whether serum, urine, or tissue assays for these proteins are of clinical relevance. The first specific aim of this proposal concerns the production of monoclonal antibodies to these proteins through the immunization of mice with protein spots cut from two-dimensional electrophoresis gels, a recently developed technique. The second aim deals with the use of these antibodies to help isolate and characterize the proteins as to molecular weight, subunit composition, glycosylation, cellular localization, and tissue specificity. These studies will use two-dimensional electrophoresis, affinity chromatography, and immunohistochemical staining. The third specific aim deals with the evaluation of three established renal cell carcinoma-derived cell lines as model systems. Quantitative computer-aided pattern analysis will be used to compare two-dimensional gels of the cultured cells and tumor tissue, and immunologic methods will also be used to confirm the presence or absence of the carcinoma-associated proteins in the cultured cells. Specific aim four will be to determine by immunoassay if any of the carcinoma-associated proteins or circulating antibodies to these proteins are present in the serum of patients with this tumor. If so, preliminary studies will be initiated to determine the potential clinical usefulness for the detection, staging, or typing of renal tumors. The studies outlined here should provide fundamental information about the biochemistry, cell biology, and clinical chemistry of a group of proteins associated with renal cell carcinoma and provide the necessary tools and information to pursue one or several of these proteins in greater detail in the future. Moreover, should clinically useful markers be developed, this would point the way towards the use of these same methods in other tumor systems. (1)